Characterization of Botrytis cinerea from table grapes in Chile using RAPD-PCR

Abstract

Random amplified polymorphic DNA (RAPD) analysis was performed on 29 isolates of Botrytis cinerea Pers. ex Fr. isolated from table grapes (Vitis vinifera L.) and other crops in Chile with 29 decaprimers. No single primer was found to differentiate either the host or the geographical origin of each of the B. cinerea isolates tested. The DNA profiles obtained, particularly with primers OPA4 and OPA11, distinguished isolates of B. cinerea from other epiphyte fungi found on table grapes, including Alternaria alternata, Aspergillus niger; Cladosporium herbarum, Epiccocum nigrum; Rhizopus stolonifer, a Penicillium sp., and yeasts (Cryptococcus laurentii, Rhodotorula glutinis, and Saccharomyces cerevisiae). Regardless of host origin, primers OPA4 and OPA11 amplified a strong fragment of 1.2 kilobases (kb) and two fragments of 1.10 and 0.7 kb, respectively. These DNA fragments were obtained even when only one conidium of B. cinerea was in the test sample. Three main groups were clearly defined based on the genetic similarities found in additional RAPD analysis with 19 arbitrary decaprimers and 15 selected isolates of B. cinerea. The overall similarity coefficients (SC) between the groups obtained ranged from 0.326 to 0.891. Interestingly, all isolates from table grapes were included in group I(SC: 0.761 to 0.826), isolates from apple and tomato were in group II (SC: 0.739 to 0.848), while isolates from blueberry were either in group I (SC: 0.804) or III (SC: 0.673). Consequently, the genetic variability determined by RAPD analysis among these B. cinerea isolates suggested a possible host:pathogen relationship. However, further research is needed to clarify its pathological significance.